2×PCR Master Mix (without Dye)
PCR Master Mix is a kind of conventional PCR premixed solution which is ready to use, including Taq DNA Polymerase, dNTP mix MgCl2 and optimized buffer. During the reaction, only the primer and template can be added for amplification, which greatly simplifies the operation steps of experiment. This product contains excellent stabilizers and can be stored for 3 months at 4℃. The PCR product have 3′-dA protrusion and can be easily cloned into T vector.
Storage Conditions
The product should be stored at -25℃~ -15℃ for two years.
Specifications
|
Fidelity (vs.Taq) |
1× |
|
Hot Start |
No |
|
Overhang |
3′-A |
|
Polymerase |
Taq DNA Polymerase |
|
Reaction Format |
SuperMix or Master Mix |
|
Reaction Speed |
Standard |
|
Product Type |
PCR Master Mix (2×) |
Instructions
1. Reaction System
|
Components |
Size (μL) |
|
Template DNA |
Suitable |
|
Primer 1 (10 μmol/L) |
2 |
|
Primer 2 (10 μmol/L) |
2 |
|
PCR Master Mix |
25 |
|
ddH2O |
To 50 |
2. Amplification Protocol
|
Cycle steps |
Temperature (°C) |
Time |
Cycles |
|
Pre-denaturation |
94 ℃ |
5mins |
1 |
|
Denaturation |
94 ℃ |
30 sec |
35 |
|
Annealing |
50-60 ℃ |
30 sec |
|
|
Extension |
72 ℃ |
30-60sec/kb |
|
|
Final Extension |
72 ℃ |
10mins |
1 |
Notes:
1) Template usage: 50-200 ng genomic DNA; 0.1- 10 ng plasmid DNA.
2) Mg2+ concentration: This product contains 3 mM of MgCl2 suitable for most PCR reactions.
3) Annealing temperature: Please refer to the theoretical Tm value of Primers. The annealing temperature can be set to 2-5 ℃ lower than the theoretical value of the primer.
4) Extension time: For molecular identification, 30 sec/kb is recommended. For gene cloning, 60 sec/kb is recommended.
Notes
1. For your safety and health, please wear lab coats and disposable gloves for operation.
2. For research use only!
